Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning
Journal of the American Chemical Society
2014
E. Barbet-Massin,
A.J. Pell,
J.S. Retel,
L.B. Andreas,
Kristaps Jaudzems,
W.T. Franks,
A.J. Nieuwkoop,
M. Hiller,
P. Guerry,
V. Higman,
A. Bertarello,
M.J. Knight,
M. Felletti,
T. Le Marchand,
S. Kotelovica,
I. Akpjana,
K. Tars,
M. Stoppini,
V. Bellotti,
M. Bolognesi,
S. Ricagno,
J.J. Chou,
R.G. Griffin,
H. Oschkinat,
A. Lesage,
L Emsley,
T. Herrmann,
G. Pintacuda
Using a set of six (1)H-detected triple-resonance NMR experiments, we establish a method for sequence-specific backbone resonance assignment of magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra of 5-30 kDa proteins. The approach relies on perdeuteration, amide (2)H/(1)H exchange, high magnetic fields, and high-spinning frequencies (ωr/2π ≥ 60 kHz) and yields high-quality NMR data, enabling the use of automated analysis. The method is validated with five examples of proteins in different condensed states, including two microcrystalline proteins, a sedimented virus capsid, and two membrane-embedded systems. In comparison to contemporary (13)C/(15)N-based methods, this approach facilitates and accelerates the MAS NMR assignment process, shortening the spectral acquisition times and enabling the use of unsupervised state-of-the-art computational data analysis protocols originally developed for solution NMR.
DOI
10.1021/ja507382j
Hipersaite
http://pubs.acs.org/doi/abs/10.1021/ja507382j
Barbet-Massin, E., Pell, A., Retel, J., Andreas, L., Jaudzems, K., Franks, W., Nieuwkoop, A., Hiller, M., Guerry, P., Higman, V., Bertarello, A., Knight, M., Felletti, M., Le Marchand, T., Kotelovica, S., Akpjana, I., Tars, K., Stoppini, M., Bellotti, V., Bolognesi, M., Ricagno, S., Chou, J., Griffin, R., Oschkinat, H., Lesage, A., Emsley, L., Herrmann, T., Pintacuda, G. Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning. Journal of the American Chemical Society, 2014, Vol.136, Iss.35, 12489.-12497.lpp. ISSN 0002-7863. e-ISSN 1520-5126. Pieejams: doi:10.1021/ja507382j
Publikācijas valoda
English (en)